Plant and Fungal Systematics 65(1): 76–94, 2020 ISSN 2544-7459 (print) DOI: https://doi.org/10.35535/pfsyst-2020-0004 ISSN 2657-5000 (online) Emmanuelia, a new genus of lobarioid lichen-forming fungi (Ascomycota: Peltigerales): phylogeny and synopsis of accepted species Antoine Simon1, Robert Lücking2, Bibiana Moncada3, Joel A. Mercado-Díaz4,5, Frank Bungartz6,7,8, Marcela Eugenia da Silva Cáceres9, Emerson Luiz Gumboski10, Suzana Maria de Azevedo Martins11, Adriano A. Spielmann12, Dinah Parker13,14 & Bernard Goffinet13* Abstract. The former family Lobariaceae, now included in Peltigeraceae as subfamily Article info Lobarioideae, has undergone substantial changes in its generic classification in recent Received: 30 Jan. 2020 years, based on phylogenetic inferences highlighting the polyphyly of the speciose genera Revision received: 11 Mar. 2020 Lobaria, Pseudocyphellaria and Sticta. Here we introduce the new genus Emmanuelia, Accepted: 16 Mar. 2020 named in honor of Prof. Emmanuël Sérusiaux for his extensive work on the Peltigerales. Published: 2 Jun. 2020 Emmanuelia currently comprises twelve species. It is superficially similar to the lobarioid Associate Editor genus Ricasolia, but differs by its apothecia, rimmed by overarching and often crenulate to Nicolas Magain lobulate margins, with the parathecium (proper excipulum) and the amphithecium (thalline excipulum formed by the thallus cortex) apically separated and of a different structure. Also, ascospore dimensions and shape differ between the two genera, with the ascospores of Emmanuelia being longer and narrower. Molecular phylogenetic analyses using DNA nucleotide sequences of the internal transcribed spacer region (ITS) and the small subu- nit of mitochondrial ribosomal DNA (mtSSU) confirm that Emmanuelia belongs to the Lobaria s.lat. clade and forms a monophyletic group sister to the lineage consisting of Dendriscosticta, Lobariella and Yoshimuriella. None of the available generic names of lobarioid lichens can be applied to this group, and consequently a new name is proposed for this new genus, which is typified with E. ravenelii comb. nov. Eleven other species are transferred to Emmanuelia: E. americana comb. nov., E. conformis comb. nov., E. cuprea comb. nov., E. elaeodes comb. nov., E. erosa comb. nov., E. excisa comb. nov., E. lob- ulifera comb. nov., E. ornata comb. nov., E. patinifera comb. nov., E. pseudolivacea comb. nov. and E. tenuis comb. nov. The genus is represented in North America by three species, including E. lobulifera, which is resurrected from synonymy with E. (Lobaria) tenuis, a South American species, and E. ornata, whose populations were previously treated under E. (Lobaria) ravenelii. Key words: Brazil, Dendriscocaulon, Lobarioideae, Neotropics, Peltigeraceae, Ricasolia, taxonomy, USA 1 Evolution and Conservation Biology, InBIOS Research Center, Uni- 9 Departamento de Biociências, Universidade Federal de Sergipe, CEP: versity of Liège, Quartier Vallée 1, Chemin de la Vallée 4, B-4000 49500-000, Itabaiana, Sergipe, Brazil Liège, Belgium 10 Universidade da Região de Joinville, Departamento de Ciências 2 Botanischer Garten und Botanisches Museum, Freie Universität Berlin, Biológicas, Laboratório de Liquenologia, Rua: Paulo Malschitzki, Germany n° 10 – Zona Industrial, CEP: 89219-710 – Joinville, SC, Brazil 3 Universidad Distrital Francisco José de Caldas, Torre de Laboratorios, 11 Fundação Zoobotânica do Rio Grande do Sul, Museu de Ciências Herbario, Bogotá, 11021, Colombia Naturais, Porto Alegre, RS, Brasil 4 Field Museum, Science and Education, Chicago, IL 60605, USA 12 Laboratório de Botânica / Liquenologia, Instituto de Biociências, Uni- 5 Committee on Evolutionary Biology, University of Chicago, 1025 E. versidade Federal de Mato Grosso do Sul, CEP 79070-900, Campo 57th Street, Chicago, IL 60637, USA Grande – MS, Brazil 6 Biodiversity Integration Knowledge Center, Arizona State University, 13 Department of Ecology and Evolutionary Biology, University of PO Box 874108, Arizona State University, Tempe, AZ 85287-4108, USA Connecticut, Storrs, CT 06269-3043, USA 7 Instituto Nacional de Biodiversidad (INABIO), Quito, Ecuador 14 Department of Plant and Environmental Sciences, University of 8 Charles Darwin Foundation for the Galapagos Islands (CDF), Galá- Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark pagos, Ecuador * Corresponding author e-mail: [email protected] © 2020 W. Szafer Institute of Botany, Polish Academy of Sciences. This is an Open Access article distributed under the terms of the Creative Commons Attribution License CC BY 4.0 (http://creativecommons.org/licenses/by/4.0/) A. Simon et al.: Emmanuelia, a new genus of lobarioid lichen-forming fungi 77 Introduction Material and methods Recent years have witnessed major systematic rearrange- Taxon sampling and phenotypic characterization ments of lobarioid lichens. In less than a decade, the The present study is based on detailed examination of number of genera circumscribed in this lineage of con- lichen specimens provided by NY (William and Lynda spicuous macrolichens was multiplied by four. Essentially, Steere Herbarium, New York, USA) and numerous freshly the three long-established genera (Lobaria, Pseudocy- collected specimens from fieldtrips to Brazil, the Carib- phellaria, Sticta), which were diagnosed by single mor- bean Islands and Galapagos Islands. Thirty-four represent- phological features (the presence/absence of cyphellae or atives of the taxonomic group of interest were selected pseudocyphellae), did not survive the advent of molecular based on preliminary phylogenetic analysis of the internal phylogenetics and were partitioned into twelve genera transcribed spacer region (ITS), along with ten speci- (Galloway & Elix 2013; Moncada et al. 2013; Galloway mens from related genera. Morphological features were 2015; McCune et al. 2014). As an example, moon lichens, observed at various laboratories, using various dissecting which were characterized by the presence of crater-like microscopes (Olympus SZ60, Leica Zoom 2000) and pores on the lower cortex, are no longer considered as compound microscopes (Olympus BX53, Nikon Eclipse a monophyletic group under the genus name Sticta, as 80i, Zeiss Axioskop). Secondary chemistry was investi- this trait evolved in two unrelated lineages. Thus, the gated through thin-layer chromatography (TLC) using sol- genus Dendriscosticta, more closely related to Lobaria, vent G and following the protocol by Orange et al. (2001). was introduced to accommodate the additional lineage Detailed descriptions were provided for the generic type (Moncada et al. 2013). of the newly introduced genus (E. ravenelii) and for the In addition to the profound changes in generic con- resurrected species (E. lobulifera). For all other species cepts, lobarioid lichens were not spared from a recent we added short diagnostic descriptions. systematic revision at the family level: in a recent study by Kraichak et al. (2018), under a temporal-banding DNA extraction, amplification, and sequencing proposal (Avise & Johns 1999; Kraichak et al. 2017), the authors proposed treating the families Lobariaceae Genomic DNA was isolated using Nucleospin Plant II and Nephromataceae as synonyms of Peltigeraceae. Midi kits (Macherey-Nagel, Bethlehem, Pennsylvania, While the mechanistic approach of temporal banding USA), following the manufacturer’s guidelines or fol- classifications has been criticized, the broad agreement lowing the protocol by Cubero et al. 1999. We amplified in morphological, anatomical and chemical features and sequenced the internal transcribed spacer region (ITS) and the absence of a clear diagnostic character for using primers ITS1F (Gardes & Bruns 1993) and ITS4 each of the three previously separated families jus- (White et al. 1990), and the small subunit of mitochon- tifies this revised classification (Lücking 2019). As drial ribosomal DNA (mtSSU) using primers SSU1 and a consequence, lobarioid lichens, long treated as Lobar- SSU3R (Zoller et al. 1999). Standard PCR protocols were iaceae, are now recognized as members of the sub- carried out using GoTaq Green Master Mix (Promega, family Lobarioideae within Peltigeraceae (Lumbsch Madison, Wisconsin, USA), following the manufacturer’s & Leavitt 2019). guidelines. The thermal cycling parameters were set as In the present study, yet another new genus, Emma- follows: 94°C for 3 min, followed by 35–40 cycles of nuelia, is erected to accommodate a group of lobarioid 94°C for 1 min, 52°C for 1 min and 70°C for 1 min, lichens that cannot be placed in any of the existing gen- with a final extension of 70°C for 10 min. The quality era. The species of interest belong to a lineage mostly and size of the amplicons were visually checked on a 1% restricted to the Neotropics and the southeastern United w/v agarose gel stained by SYBR Safe DNA Gel Stain States, and were previously treated as members of the (Thermo Fisher Scientific Inc., Waltham, Massachusetts, genus Lobaria and subsequently considered part of USA). Amplicons were cleaned using the ExoSAP-IT Ricasolia (Moncada et al. 2013; Käffer et al. 2016; Leh- protocol (USB Corporation, Cleveland, Ohio, USA) nen et al. 2017; Etayo et al. 2018). Yoshimura (1998) and sequenced on a ABI Prism 3100 Genetic Analyzer treated the South American taxa as L. quercizans group, (Applied Biosystems, Foster City, California, USA), or also implying a close relationship to Ricasolia. This by Macrogen, Inc. (Seoul, South Korea). The forward and group of lichens includes,